pUC19L

Linearized version of pUC19 for use with the GeneArt® Seamless Cloning and Assembly Kit.

Sequence Author: Thermo Fisher (Invitrogen)

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End (2659) BanII - SacI (2653) Eco53kI (2651) ApoI - EcoRI (2643) PluTI (2486) SfoI (2484) NarI (2483) KasI (2482) BstAPI (2432) NdeI (2431) PfoI (2293) EcoO109I (2236) AatII (2182) ZraI (2180) SspI (2064) XmnI (1859) ScaI (1740) TsoI (1659) NmeAIII (1408) BsrFI (1340) BpmI (1330) Start (0) SphI (6) HindIII (8) BspQI - SapI (251) AflIII - PciI (367) BseYI (671) PspFI (675) AlwNI (783) AhdI (1260) BsaI (1321) pUC19L 2659 bp
End  (2659)
0 sites
BanII  (2653)
1 site
G R G C Y C C Y C G R G

Sticky ends from different BanII sites may not be compatible.
SacI  (2653)
1 site
G A G C T C C T C G A G
Eco53kI  (2651)
1 site
G A G C T C C T C G A G
ApoI  (2643)
1 site
R A A T T Y Y T T A A R

ApoI is typically used at 50°C, but is 50% active at 37°C.
EcoRI  (2643)
1 site
G A A T T C C T T A A G
PluTI  (2486)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the PluTI recognition sequence.
SfoI  (2484)
1 site
G G C G C C C C G C G G
NarI  (2483)
1 site
G G C G C C C C G C G G

Efficient cleavage requires at least two copies of the NarI recognition sequence.
KasI  (2482)
1 site
G G C G C C C C G C G G
BstAPI  (2432)
1 site
G C A N N N N N T G C C G T N N N N N A C G

Sticky ends from different BstAPI sites may not be compatible.
NdeI  (2431)
1 site
C A T A T G G T A T A C

Prolonged incubation with NdeI may lead to removal of additional nucleotides.
PfoI  (2293)
1 site
T C C N G G A A G G N C C T

Sticky ends from different PfoI sites may not be compatible.
EcoO109I  (2236)
1 site
R G G N C C Y Y C C N G G R

Sticky ends from different EcoO109I sites may not be compatible.
AatII  (2182)
1 site
G A C G T C C T G C A G
ZraI  (2180)
1 site
G A C G T C C T G C A G
SspI  (2064)
1 site
A A T A T T T T A T A A
XmnI  (1859)
1 site
G A A N N N N T T C C T T N N N N A A G
ScaI  (1740)
1 site
A G T A C T T C A T G A
TsoI  (1659)
1 site
T A R C C A ( N ) 9 N N A T Y G G T ( N ) 9

Sticky ends from different TsoI sites may not be compatible.
After cleavage, TsoI can remain bound to DNA and alter its electrophoretic mobility.
For full activity, add fresh S-adenosylmethionine (SAM).
NmeAIII  (1408)
1 site
G C C G A G ( N ) 18-19 N N C G G C T C ( N ) 18-19

Efficient cleavage requires at least two copies of the NmeAIII recognition sequence.
Sticky ends from different NmeAIII sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
BsrFI  (1340)
1 site
R C C G G Y Y G G C C R

Cleavage may be enhanced when more than one copy of the BsrFI recognition sequence is present.
After cleavage, BsrFI can remain bound to DNA and alter its electrophoretic mobility.
BpmI  (1330)
1 site
C T G G A G ( N ) 14 N N G A C C T C ( N ) 14

Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
Start  (0)
0 sites
SphI  (6)
1 site
G C A T G C C G T A C G
HindIII  (8)
1 site
A A G C T T T T C G A A
BspQI  (251)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different BspQI sites may not be compatible.
SapI  (251)
1 site
G C T C T T C N C G A G A A G N N N N

Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
AflIII  (367)
1 site
A C R Y G T T G Y R C A

Sticky ends from different AflIII sites may not be compatible.
PciI  (367)
1 site
A C A T G T T G T A C A

PciI is inhibited by nonionic detergents.
BseYI  (671)
1 site
C C C A G C G G G T C G

After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
PspFI  (675)
1 site
C C C A G C G G G T C G
AlwNI  (783)
1 site
C A G N N N C T G G T C N N N G A C

Sticky ends from different AlwNI sites may not be compatible.
AhdI  (1260)
1 site
G A C N N N N N G T C C T G N N N N N C A G

The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
BsaI  (1321)
1 site
G G T C T C N C C A G A G N ( N ) 4

Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
AmpR
1187 .. 2047  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   1187 .. 1978  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1187 .. 2047  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   1979 .. 2047  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1187 .. 2047  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
428 .. 1016  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
428 .. 1016  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
AmpR promoter
2048 .. 2152  =  105 bp
AmpR promoter
2048 .. 2152  =  105 bp
lac promoter
74 .. 104  =  31 bp
3 segments
   Segment 3:  -10  
   74 .. 80  =  7 bp
promoter for the E. coli lac operon
lac promoter
74 .. 104  =  31 bp
3 segments
   Segment 2:  
   81 .. 98  =  18 bp
promoter for the E. coli lac operon
lac promoter
74 .. 104  =  31 bp
3 segments
   Segment 1:  -35  
   99 .. 104  =  6 bp
promoter for the E. coli lac operon
lac promoter
74 .. 104  =  31 bp
3 segments
promoter for the E. coli lac operon
M13 rev
26 .. 42  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
26 .. 42  =  17 bp
common sequencing primer, one of multiple similar variants
lac operator
50 .. 66  =  17 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
50 .. 66  =  17 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
M13 fwd
2626 .. 2642  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
2626 .. 2642  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  1317 .. 1583  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  2433 .. 2657  =  225 bp
ORF:  75 amino acids  =  8.8 kDa
ORF:  1187 .. 2047  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  2393 .. 2659  =  267 bp
ORF:  88 amino acids  =  10.1 kDa  (no start codon)
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Individual Sequences & Maps

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